荷叶碱抑制肝脏脂质沉积改善II型糖尿病及其分子机制研究

从植物化学物中寻找具有防治II型糖尿病（T2DM）的先导化合物是新药开发领域研究的热点。荷叶碱是一种阿朴啡类生物碱具有许多药理学功效。前期发现荷叶碱可抑制小鼠肝脏脂质沉积、促进核受体PPARα及其共激活子PGC-1α的活性而缓解T2DM，但其作用机制不清楚。本项目拟以脂质沉积肝细胞和T2DM小鼠为模型，筛选出荷叶碱有效处理浓度和方式；通过对血清、肝脏及脂肪组织的生化及病理分析，探明其对T2DM小鼠脂质沉积、糖脂代谢紊乱及胰岛素抵抗的缓解作用；通过质谱、光谱、滴定量热、RT-qPCR、siRNA、免疫组化/共沉淀等阐明荷叶碱对PPARα和PGC-1α的激活机制，揭示二者互作的热/动力学规律；利用基因敲除鼠阐明荷叶碱对T2DM肝脏脂质沉积的缓解是否依赖于PPARα-PGC-1α通路的激活，提出其防控T2DM的可能机制。本项目不仅能明晰荷叶碱的新功效，也为新型抗T2DM药物的研发提供新思路。

It is a research hotspot that the search of phytochemicals as lead compounds for the prevention and treatment of type 2 diabetes mellitus (T2DM) in the field of new drug development. Nuciferine is an aporphine alkaloid with many pharmacological activities. Our preliminary data found that nuciferine can ameliorate T2DM by inhibiting lipid deposition in the liver of mice, promoting the activity of nuclear receptor PPARα and its co-activator PGC-1α, but the mechanism of action is unclear. In this project, lipid deposition hepatocytes and T2DM mice are used as models to screen out the effective treatment concentrations and methods of nuciferine. Through biochemical and pathological analysis of serum, liver and adipose tissues, the alleviating effects of T2DM mice on lipid deposition, glucose, and lipid metabolism disorders and insulin resistance will be explored. The activation mechanism of nuciferine based on PPARα and PGC-1α is verified by mass spectrometry, spectrum, isothermal titration calorimetry, RT-qPCR, siRNA, immunohistochemistry/co-precipitation, etc. Furthermore, the regularities of thermodynamics and dynamics of the interaction between PPARα and PGC-1α will be revealed. Moreover, the knockout mice will be used to clarify whether the alleviation of hepatic lipid deposition by nuciferine on T2DM depending on the activation of PPARα/PGC-1α signaling pathway. Together, the possible anti-T2DM mechanism of nuciferine will be proposed according to the above elaboration. This study can not only clarify novel pharmacological activities of nuciferine but also provide new ideas for the research and development of new natural drugs for anti-T2DM.